Journal of Fish Biology (2001) 59 (Supplement A), 190–208 doi:10.1006/jfbi.2001.1766, available online at http://www.idealibrary.com on Anthropogenic changes in the freshwater fish fauna of Italy, with reference to the central region and Barbus graellsii,a newly established alien species of Iberian origin P. G. B*‡ V. K† *Department of Zoology, University Federico II, Via Mezzocannone 8, 80134 Naples, Italy and †Department of Animal and Human Biology, University ‘ La Sapienza ’, Viale dell’Universita` 32, 00100 Rome, Italy In a survey of the west-central Italian rivers Ombrone, Fiora, Albegna and Bruna, among established exotic species, the Iberian barbel Barbus graellsii was recorded in Italian fresh waters for the first time. Morphological identification was supported by comparison of cytochrome b sequences with those from related barbel species. Other exotics of particular note were Barbus barbus, Pseudorasbora parva and Leuciscus cephalus, together with the Padano-Venetian Chondrostoma genei and Padogobius bonelli. Native species still present included Leuciscus lucumonis, Telestes muticellus, Rutilus rubilio and Padogobius nigricans, but were now more restricted to upper reaches and smaller watercourses. The deleterious effect of alien species on native forms is discussed with particular reference to probable competition between C. genei and L. lucumonis, and between the two gobies. The checklist of species introduced to the fresh waters of Italy is now updated to 34. For the Italian freshwater fish fauna in general, the conservation status of three native species (Acipenser naccari, Salmo marmoratus and Knipowitschia punctatissima) has been improved but five species are believed to be at risk (Salmo carpio, L. lucumonis, Scardinius scardafa, Gobio benacensis and P. nigricans) and four anadromous species (Petromyzon marinus, Lampetra fluviatilis, Acipenser sturio and Huso huso) no longer breed in Italian fresh waters. The processes of change in the com- position of the Italian freshwater fish fauna as a whole may be summarized as successively ‘ padanization ’, ‘ danubization ’ and now ‘ globalization ’. 2001 The Fisheries Society of the British Isles Key words: freshwater fishes; Italy; alien species; translocation; molecular taxonomy; Barbus graellsii. INTRODUCTION Italy has been a country long interested in the introduction of alien freshwater fishes. The first of these probably occurred during the Roman period or earlier, and several species traditionally considered as native could be of non-native origin in view of their bioecological and biogeographical features (Bianco, 1998a). At present the major centres providing species for stocking public waters are located in northern Italy and supply ‘ white fishes ’ (a mixture of species, mostly chubs and barbels, but with several others included accidentally) (Bianco, 1990a, 1998a; Melotti, 1994). It has been calculated that, in recent times, these farms have imported c. 100 t every week of such fishes from outside Italy, mostly eastern Europe (Ielardi, 1998). This fishes are bought and officially released by the provincial authorities in every river. Following these practices, such species ‡Author to whom correspondence should be addressed. Tel.: +39 81 790 3349; fax: +39 81 790 3342; email: [email protected] 190 0022–1112/01/59A190+19 $35.00/0 2001 The Fisheries Society of the British Isles 191 as the European Barbus barbus (L.), recorded in Italy in 1994 (Bianco, 1995a) and the Asiatic Pseudorasbora parva (Temminck & Schlegel), found in Italy in 1987 (Sala & Spampanato, 1991), have reached in a few years an almost pan-Italian distribution. The introduction and establishment is now reported of another alien species, the Iberian barbel Barbus graellsii Steindachner (Bianco, 1998b), identified by both morphological and sequencing criteria. In the last two decades such methodology, driven by molecular biology, has been successfully applied to solve several systematic problems with European freshwater fishes (Durand et al., 2000). This species has been found during surveys of an area including the rivers Bruna, Ombrone, Albegna and Fiora from Grosseto and Siena Provinces in western central Italy. The Iberian barbel was found with the Danubian B. barbus and the native Barbus tyberinus Bonaparte. The Provincial Administration of Grosseto has intensively stocked the water bodies of its territory with white fishes for at least 20–25 years. In 1996, for instance, 1500 kg of barbels were released at 11 sites on the Ombrone, Fiora, Albegna and Orcia, with material from several fish farms, with extensive stockings of ‘ non-barbel ’ species (pike Esox lucus L., trout Salmo trutta L., chub Leuciscus cephalus (L.), carp Cyprinus carpio L., tench Tinca tinca (L.)) (G. Giarola, pers. comm.). The Iberian barbel is thought to have been introduced in this way. About three other alien species have become established in the last five years. Such introductions have resulted in a radical change in the composition of the Italian freshwater fish fauna with the local extinction or decline of native populations. A balance between alien and native fishes in many water bodies is still to be reached. MATERIALS AND METHODS The Rivers Ombrone, Fiora, Albegna and Bruna were surveyed on several occasions during 1994–1995 (Bianco, 1995b) and also in 2000 as part of a provincial programme for the reintroduction of the otter Lutra lutra (L.). About 25 different localities were surveyed at least twice, in spring-summer and autumn-winter respectively. Fishes were collected by electrofishing. Each specimen was measured (total length, LT), weighed, some scales collected for age determination, and then released. Only a few specimens, especially those of the alien species, were killed for morphological, biological and genetic analyses. Two small specimens of B. tyberinus and of the putative Iberian Barbus, and three specimens of B. barbus, were fixed immediately in ethyl alcohol for DNA analysis. The mitochondrial cytochrome b gene (cytb) was chosen because of its usefulness for distinguishing morphologically similar species of cyprinids (Zardoya & Doadrio, 1998). Further details of extraction methods and sequencing for species identification are given in Appendix I. For the morphological comparisons preserved specimens were used of several Iberian Barbus. The total quantity of fish collected in the three basins permitted the description of local communities by category ‘ native ’, ‘ translocated ’ (to the study area from northern Italy), and ‘ exotic ’ (from outside Italy), number of species, number of individuals, and biomass. Twelve sites were selected to show the influence of alien species on native assemblages in different biotopes, namely, six sites from the middle and low courses (sites 1, 2, 3 and 10, 11, 12, Table I) of main rivers, and six from the upper reaches or brooks and rivulets (sites 3, 4, 5 and 7, 8, 9, Table I). Since the species Leuciscus cephalus (L.) in the study has originated from three different sources (native, transplanted from northern Italy, and introduced from the Danube area), this species has been included in each T I. Twelve selected collecting localities within the study area and number of fish species belonging to native, Padano-Venetian (Padanic) and extra-Italian (exotic) categories. Barbus graellsii was found in localities 1, 2 and 3 Collecting localities Origins 1. River 2. River. 3. River 4. Lanzo 5. Upper River 6. Upper River Ombrone, Albegna, Albegna, Brook, Albegna, Fiora, Grosseto Albinia Canino Cinigiano Arcidosso Fiora Native 4 4 4 7 5 4 Padanic 6 3 2 3 1 1 Exotic 5 6 3 0 0 0 Collecting localities Origins 7. La Gonna 8. Upper River 9. Upper River 10. River 11. River 12. River Brook, Farma, Merse, Ombrone, Ombrone, Merse, Monticiano Roccastrada San Galgano Buonconvento Montalcino S. Lorenzo Native 6 7 6 4 5 5 Padanic 2 5 5 7 7 8 Exotic 1 0 2 3 4 6 193 category and the number of fish and the biomass divided by two. The chub populations found in lower courses of rivers, together with the Iberian barbel (sites 1, 2 and 3, Table I), have been considered to be all of Danubian origins since they have modally eight branched rays in the anal fin which is typical for Danubian chub. RESULTS A list of species collected in the study area with their areas of original distribution and populations trends is given in Table II. The following notes concern the Iberian barbel, other exotic species, Padano-Venetian species established in the area, and native species. IBERIAN BARBEL Three species of Barbus were found in the waters surveyed, the introduced Danubian B. barbus, the native B. tyberinus, and a third species which is now identified as the Iberian barbel Barbus graellsii Steindachner, an endemic species of the River Ebro in Spain, and belonging to the ‘ Messinobarbus ’ group characterized by the prominent nuptial tubercles on the snout in adult males (Bianco, 1998b)(Fig. 1). A comparisons of morphological features of several Barbus species (Table III) supports identification of the Iberian barbel and the present material corresponds well with the description and illustration given by Doadrio et al. (1991) for B. graellsii. It is believed to have been introduced to the study area by stocking of fish from northern Italian distributors. Comparison of cytochrome b sequences derived from the putative Iberian barbel in the present work with GenBank sequences for several Barbus taxa (Table III), showed a highest identity score (99%) with that of Barbus graellsii (Accession Number AF045973) All other Iberian species examined had smaller identity scores, ranging from 95 to 97%, and the lowest values were those found in comparisons with B. tyberinus and B. barbus (89% each). Among the three Barbus species found at the study sites, B. graellsii cannot be misidentified because of its uniformly silvery colouration, larger scales and ink black peritoneal membrane, in contrast to the greyish peritoneum in the other species. Gill rakers of B. graellsii are also much more numerous than in B. barbus and B. tyberinus. Another diagnostic character is the presence of large nuptial tubercles on the snout of adult males (Fig.